Anti-Calreticulin antibody (ab4)
- Product nameAnti-Calreticulin antibodySee all Calreticulin primary antibodies ...
- DescriptionRabbit polyclonal to Calreticulin
- SpecificityThis antibody is specific for calreticulin.
- Tested applicationsICC/IF, IHC-P, WB, IP, ICC more details
- Species reactivityReacts with: Mouse, Rat, Chicken, Hamster, Cow, Human, Zebrafish, African Green Monkey
Fusion protein, containing amino acids 154-347 of Mouse Calreticulin.
- Positive control
- IC: COS7 and HCT29 cells WB: H4IIE cell lysate, NBL-1 bovine renal epithelial cell lysate
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.05% Sodium Azide
- PurityWhole antiserum
- Clonality Polyclonal
- Research Areas
Our Abpromise guarantee covers the use of ab4 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||IHC-P: 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Incubate primary antibody for 1 hour.|
|WB||WB: 1/1000. Detects a band of approximately 55-60 kDa (predicted molecular weight: 48 kDa).|
- FunctionMolecular calcium-binding chaperone promoting folding, oligomeric assembly and quality control in the ER via the calreticulin/calnexin cycle. This lectin interacts transiently with almost all of the monoglucosylated glycoproteins that are synthesized in the ER. Interacts with the DNA-binding domain of NR3C1 and mediates its nuclear export.
- Sequence similaritiesBelongs to the calreticulin family.
- DomainCan be divided into a N-terminal globular domain, a proline-rich P-domain forming an elongated arm-like structure and a C-terminal acidic domain. The P-domain binds one molecule of calcium with high affinity, whereas the acidic C-domain binds multiple calcium ions with low affinity.
The interaction with glycans occurs through a binding site in the globular lectin domain.
The zinc binding sites are localized to the N-domain.
Associates with PDIA3 through the tip of the extended arm formed by the P-domain.
- Cellular localizationEndoplasmic reticulum lumen. Cytoplasm > cytosol. Secreted > extracellular space > extracellular matrix. Cell surface. Also found in cell surface (T cells), cytosol and extracellular matrix. Associated with the lytic granules in the cytolytic T-lymphocytes.
- Entrez Gene: 281036 Cow
- Entrez Gene: 811 Human
- Entrez Gene: 12317 Mouse
- Entrez Gene: 64202 Rat
- Entrez Gene: 30248 Zebrafish
- Omim: 109091 Human
- SwissProt: P52193 Cow
- SwissProt: P27797 Human
- SwissProt: P14211 Mouse
- SwissProt: Q3TVD2 Mouse
- SwissProt: P18418 Rat
- SwissProt: Q6NV30 Zebrafish
- SwissProt: Q6PE26 Zebrafish
- SwissProt: Q9PUC1 Zebrafish
- Unigene: 515162 Human
- Unigene: 1971 Mouse
- Unigene: 467043 Mouse
- Unigene: 974 Rat
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Anti-Calreticulin antibody images
All lanes : Anti-Calreticulin antibody (ab4) at 1/1000 dilution
Lane 1 : Chicken lymphoid cell line lysate
Lane 2 : Chicken lymphoid cell line lysate
Lysates/proteins at 1000000 cells per lane.
Polyclonal HRP-conjugate Goat anti-rabbit IgG at 1/2000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 48 kDa
Observed band size : 55 kDa (why is the actual band size different from the predicted?)
Additional bands at : 58 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 30 seconds
This image is courtesy of an anonymous Abreview
IF staining was performed on HCT15 colon cancer cells using a 1:50 dilution of the primary antibody followed by hybridisation with a 1:100 dilution of the secondary anti-rabbit antibody Alexa Fluor 488 (molecular Probes) with green fluorescence. Picture from review by Karin Birkenkamp-Demtroeder submitted 20 July 2004
ICC/IF image of ab4 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab4, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
ab4 staining calreticulin in Human placenta.
Left panel: with primary antibody diluted 1:4000. Right panel: isotype control.
Sections were stained using an automated system (DAKO Autostainer Plus), at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers EDTA pH 9.0. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
References for Anti-Calreticulin antibody (ab4)
This product has been referenced in:
- Toptan T et al. Complex alternative cytoplasmic protein isoforms of the Kaposi's sarcoma-associated herpesvirus latency-associated nuclear antigen 1 generated through noncanonical translation initiation. J Virol 87:2744-55 (2013). Read more (PubMed: 23255808) »
- Li Q et al. Estradiol accelerates the effects of fluoxetine on serotonin 1A receptor signaling. Psychoneuroendocrinology 38:1145-57 (2013). WB ; Rat . Read more (PubMed: 23219224) »